Sakai-Kato Kumiko, Kinouchi Tadatoshi, Fujii Noriko, Imai Kazuhiro, Utsunomiya-Tate Naoko
Research Institute of Pharmaceutical Sciences, Musashino University, 1-1-20 Shinmachi, Nishitokyo-shi, Tokyo, 202-8585, Japan.
Amino Acids. 2009 Jan;36(1):125-9. doi: 10.1007/s00726-008-0040-y. Epub 2008 Feb 13.
D-Asp-containing proteins have been implicated in many aging-related diseases. To clarify the role of D-Asp-containing proteins in such diseases, we developed a screening system for these proteins using a D-aspartyl endopeptidase that specifically cleaves the proteins at the C-terminus. The digested proteins were detected by means of two-dimensional gel electrophoresis and identified using nano-liquid chromatography/tandem mass spectrometry. We were able to detect myelin basic protein, a known D-Asp-containing protein, in the brain tissues of mice; this indicates that our system is effective for screening D-Asp-containing proteins.
含D-天冬氨酸的蛋白质与许多衰老相关疾病有关。为了阐明含D-天冬氨酸的蛋白质在这些疾病中的作用，我们利用一种能在C端特异性切割这些蛋白质的D-天冬氨酰内肽酶，开发了一种针对这些蛋白质的筛选系统。通过二维凝胶电泳检测消化后的蛋白质，并使用纳升液相色谱/串联质谱进行鉴定。我们能够在小鼠脑组织中检测到髓鞘碱性蛋白，一种已知的含D-天冬氨酸的蛋白质；这表明我们的系统对于筛选含D-天冬氨酸的蛋白质是有效的。
